Martin A Brown

US Patent:

20030211548, Nov 13, 2003

Filed:

Jan 28, 2003

Appl. No.:

10/353791

Inventors:

Beverly Packard - Rockville MD, US
Martin Brown - Germantown MD, US
Mark Feinberg - Atlanta GA, US
Luzheng Liu - Brookline MA, US
Guido Silvestri - Decatur GA, US
Ann Chahroudi - Atlanta GA, US
Akira Komoriya - Rockville MD, US

Assignee:

Oncolmmunin, Inc.

International Classification:

C12Q001/70
G01N033/53
G01N033/567
C12Q001/37

US Classification:

435/007200, 435/023000, 435/005000

Abstract:

This invention provides a non-radioactive assay to monitor and quantify the target-cell killing activities mediated by cytotoxic T lymphocytes (CTLs). This assay is predicated on the discovery that apoptosis pathway activation and, in particular, caspase activity, provides a measure of cytotoxic effector cell activity. In one embodiment, measurement of CTL-induced caspase activation in target cells is achieved through detection of the specific cleavage of fluorogenic caspase substrates. This assay reliably detects antigen-specific CTL killing of target cells, and provides a more sensitive, more informative and safer alternative to the standard Cr-release assay most often used to quantify CTL responses. The assay can be used to study CTL-mediated killing of primary host target cells of different cell lineages, and enables the study of antigen-specific cellular immune responses in real time at the single-cell level. As such, the assay can provide a valuable tool for studies of infectious disease pathogenesis and development of new vaccines and immunotherapies.

US Patent:

20090263830, Oct 22, 2009

Filed:

Mar 3, 2009

Appl. No.:

12/396736

Inventors:

BEVERLY PACKARD - Potomac MD, US
MARTIN J. BROWN - Germantown MD, US
MARK FEINBERG - Atlanta GA, US
LUZHENG LIU - Brookline MA, US
GUIDO SILVESTRI - Decatur GA, US
ANN CHAHROUDI - Atlanta GA, US
AKIRA KOMORIYA - Potomac MD, US

Assignee:

Oncoimmunin, Inc. - Gaithersburg MD

International Classification:

G01N 33/53
C12Q 1/37
C12Q 1/02

US Classification:

435 74, 435 23, 435 29

Abstract:

This invention provides a non-radioactive assay to monitor and quantify the target-cell killing activities mediated by cytotoxic T lymphocytes (CTLs). This assay is predicated on the discovery that apoptosis pathway activation and, in particular, caspase activity, provides a measure of cytotoxic effector cell activity. In one embodiment, measurement of CTL-induced caspase activation in target cells is achieved through detection of the specific cleavage of fluorogenic caspase substrates. This assay reliably detects antigen-specific CTL killing of target cells, and provides a more sensitive, more informative and safer alternative to the standard Cr-release assay most often used to quantify CTL responses. The assay can be used to study CTL-mediated killing of primary host target cells of different cell lineages, and enables the study of antigen-specific cellular immune responses in real time at the single-cell level. As such, the assay can provide a valuable tool for studies of infectious disease pathogenesis and development of new vaccines and immunotherapies.